Polypeptide specific for mucin 1 and use thereof

Abstract

The present invention relates to a polypeptide binding to mucin 1, an isolated polynucleotide encoding same, a vector carrying the polynucleotide, and a cell including the vector. In addition, the present invention relates to a chimeric antigen receptor including the polypeptide binding to mucin 1, an isolated polynucleotide encoding the chimeric antigen receptor, a vector carrying the polynucleotide, an immune cell expressing the chimeric antigen receptor, a composition comprising same for treatment of cancer, and a method for treatment of cancer.

1 . A polypeptide binding to mucin1, the polypeptide comprising a pair of a heavy chain variable (VH) region and a light chain variable (VL) region selected from the following VH and VL regions:
 a VH region comprising complementarity-determining region (CDR) 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 43, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 46, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 54, SEQ ID NO: 55, and SEQ ID NO: 56, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 61, SEQ ID NO: 62, and SEQ ID NO: 63, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 64, SEQ ID NO: 65, and SEQ ID NO: 66, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 71, SEQ ID NO: 72, and SEQ ID NO: 73, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 74, SEQ ID NO: 75, and SEQ ID NO: 76, respectively;   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 81, SEQ ID NO: 82, and SEQ ID NO: 83, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86, respectively; and   a VH region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 91, SEQ ID NO: 92, and SEQ ID NO: 93, respectively, and a VL region comprising CDR 1, CDR 2, and CDR 3 represented by amino acid sequences of SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96, respectively.   
     
     
         2 . The polypeptide of  claim 1 , comprising a pair of a heavy chain variable (VH) region and a light chain variable (VL) region selected from the following VH and VL regions:
 a VH region comprising an amino acid sequence of SEQ ID NO: 7 and a VL region comprising an amino acid sequence of SEQ ID NO: 8;   a VH region comprising an amino acid sequence of SEQ ID NO: 17 and a VL region comprising an amino acid sequence of SEQ ID NO: 18;   a VH region comprising an amino acid sequence of SEQ ID NO: 27 and a VL region comprising an amino acid sequence of SEQ ID NO: 28;   a VH region comprising an amino acid sequence of SEQ ID NO: 37 and a VL region comprising an amino acid sequence of SEQ ID NO: 38;   a VH region comprising an amino acid sequence of SEQ ID NO: 47 and a VL region comprising an amino acid sequence of SEQ ID NO: 48;   a VH region comprising an amino acid sequence of SEQ ID NO: 57 and a VL region comprising an amino acid sequence of SEQ ID NO: 58;   a VH region comprising an amino acid sequence of SEQ ID NO: 67 and a VL region comprising an amino acid sequence of SEQ ID NO: 68;   a VH region comprising an amino acid sequence of SEQ ID NO: 77 and a VL region comprising an amino acid sequence of SEQ ID NO: 78;   a VH region comprising an amino acid sequence of SEQ ID NO: 87 and a VL region comprising an amino acid sequence of SEQ ID NO: 88; and   a VH region comprising an amino acid sequence of SEQ ID NO: 97 and a VL region comprising an amino acid sequence of SEQ ID NO: 98.   
     
     
         3 . The polypeptide of  claim 1 , wherein the polypeptide is an antibody or an antigen-binding fragment thereof. 
     
     
         4 . The polypeptide of  claim 1 , wherein the polypeptide is a single chain variable fragment (scFv), a peptibody, a fragment antigen binding (Fab), a monoclonal antibody, a bispecific antibody, a minibody, a domain antibody, a synthetic antibody, a chimeric antibody, a humanized antibody, a human antibody, or an antibody fusion protein. 
     
     
         5 . The polypeptide of  claim 1 , wherein the VH region and the VL region are linked to each other via a linker. 
     
     
         6 . The polypeptide of  claim 5 , wherein an amino acid sequence of the linker is GGGGSGGGGSGGGAS. 
     
     
         7 . The polypeptide of  claim 1 , wherein the polypeptide is a single chain variable fragment (scFv) and the scFv includes an amino acid sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 19, SEQ ID NO: 29, SEQ ID NO: 39, SEQ ID NO: 49, SEQ ID NO: 59, SEQ ID NO: 69, SEQ ID NO: 79, SEQ ID NO: 89, and SEQ ID NO: 99. 
     
     
         8 . An isolated polynucleotide encoding the polypeptide binding mucin1 of  claim 1 . 
     
     
         9 . A vector comprising the polynucleotide of  claim 8 . 
     
     
         10 . A cell comprising the vector of  claim 9 . 
     
     
         11 . A chimeric antigen receptor comprising the polypeptide binding mucin1 of  claim 1 . 
     
     
         12 . The chimeric antigen receptor of  claim 11 , comprising an extracellular domain comprising the polypeptide binding mucin1; a transmembrane domain; and an intracellular signaling domain. 
     
     
         13 . The chimeric antigen receptor of  claim 12 , wherein the extracellular domain further includes a spacer region between the polypeptide binding mucin1 and the transmembrane domain. 
     
     
         14 . The chimeric antigen receptor of  claim 13 , wherein the spacer region includes a hinge region of CD8α or CD28, or all or part of a constant region of an immunoglobulin (IgG). 
     
     
         15 . The chimeric antigen receptor of  claim 12 , wherein the transmembrane domain is a transmembrane domain of CD28 or CD8. 
     
     
         16 . The chimeric antigen receptor of  claim 12 , wherein the intracellular signaling domain is a CD3 zeta signaling domain. 
     
     
         17 . The chimeric antigen receptor of  claim 12 , further comprising at least one co-stimulatory domains. 
     
     
         18 . The chimeric antigen receptor of  claim 17 , wherein the at least one co-stimulatory domain is placed between the transmembrane domain and the intracellular signaling domain. 
     
     
         19 . The chimeric antigen receptor of  claim 17 , wherein the at least co-stimulatory domain is a signaling domain of CD28, OX-40, 4-1BB (CD137), CD27, or ICOS. 
     
     
         20 . An isolated polynucleotide encoding the chimeric antigen receptor of  claim 11 . 
     
     
         21 . A vector comprising the polynucleotide of  claim 20 . 
     
     
         22 . An immune cell expressing a chimeric antigen receptor comprising the polypeptide binding mucin1 of  claim 1 , or comprising a polynucleotide encoding the chimeric antigen receptor. 
     
     
         23 . The immune cell of  claim 22 , wherein the immune cell is a T cell, a tumor infiltrating lymphocyte (TIL), a natural killer (NK) cell, a TCR-expressing cell, a dendritic cell, or an NK-T cell. 
     
     
         24 . The immune cell of  claim 22 , wherein the immune cell is an autologous T cell or an allogenic T cell. 
     
     
         25 . A composition for preventing or treating cancer, the composition comprising the polypeptide binding mucin1 of  claim 1 ;
 an isolated polynucleotide encoding the polypeptide binding mucin1;   a vector comprising the polynucleotide encoding the polypeptide binding mucin1;   a cell comprising the polynucleotide encoding the polypeptide binding mucin1;   a chimeric antigen receptor comprising the polypeptide binding mucin1;   an isolated polynucleotide encoding the chimeric antigen receptor;   a vector comprising the polynucleotide encoding the chimeric antigen receptor; or   an immune cell comprising the polynucleotide encoding the chimeric antigen receptor, or expressing the chimeric antigen receptor.